Identification and characterization of coumestans as novel HCV NS5B polymerase inhibitors.
Identification and characterization of coumestans as novel HCV NS5B polymerase inhibitors.
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鉴定和表征香豆作为新型HCV NS5B聚合酶抑制剂。
DOI：
10.1093/nar/gkm1178
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发表时间：
2008-03
影响因子：
14.9
通讯作者：
Noel, Francois
中科院分区：
文献类型：
--
作者：
Kaushik-Basu, Neerja;Bopda-Waffo, Alain;Talele, Tanaji T.;Basu, Amartya;Costa, Paulo R. R.;da Silva, Alcides J. M.;Sarafianos, Stefan G.;Noel, Francois
丙型肝炎病毒（HCV） NS5B对病毒RNA复制至关重要，因此是开发HCV复制抑制剂的主要靶点。在这里，我们报告了一类新的HCV NS5B抑制剂的鉴定，属于植物雌激素的coumestan家族。在体外低微摩尔抑制NS5B RNA依赖RNA聚合酶（RdRp）的基础上，我们对四种具有不同a环和D环取代模式的合成coumestan类似物的抗NS5B活性进行了评价，发现它们具有良好的构效相关性。动力学表征表明，coumestans对三磷酸核苷（rNTP）底物具有非竞争性抑制模式，对核酸模板具有混合抑制模式，主要具有竞争性成分。库美斯坦与核酸底物加成实验顺序的改变影响库美斯坦抑制剂的效价。通过交联实验证实了在NS5B-RNA二元复合物形成过程中存在库米斯坦干扰。在NS5B变构位点内的coumestans分子对接，其计算的结合能与IC50值具有显著的相关性。因此，Coumestans增加了抗NS5B药物的多样性，并为结构改进和开发有效的NS5B抑制剂提供了新的支架。
The hepatitis C virus (HCV) NS5B is essential for viral RNA replication and is therefore a prime target for development of HCV replication inhibitors. Here, we report the identification of a new class of HCV NS5B inhibitors belonging to the coumestan family of phytoestrogens. Based on the in vitro NS5B RNA-dependent RNA polymerase (RdRp) inhibition in the low micromolar range by wedelolactone, a naturally occurring coumestan, we evaluated the anti-NS5B activity of four synthetic coumestan analogues bearing different patterns of substitutions in their A and D rings, and observed a good structure-activity correlation. Kinetic characterization of coumestans revealed a noncompetitive mode of inhibition with respect to nucleoside triphosphate (rNTP) substrate and a mixed mode of inhibition towards the nucleic acid template, with a major competitive component. The modified order of addition experiments with coumestans and nucleic acid substrates affected the potencies of the coumestan inhibitors. Coumestan interference at the step of NS5B–RNA binary complex formation was confirmed by cross-linking experiments. Molecular docking of coumestans within the allosteric site of NS5B yielded significant correlation between their calculated binding energies and IC50 values. Coumestans thus add to the diversifying pool of anti-NS5B agents and provide a novel scaffold for structural refinement and development of potent NS5B inhibitors.
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影响因子：
1.3
作者：
Cornberg, M;Hüppe, D;Manns, MP
通讯作者：
Manns, MP
影响因子：
5.4
作者：
Ferrari, E;Wright-Minogue, J;Hong, Z
通讯作者：
Hong, Z
影响因子：
4.8
作者：
Gu, BH;Johnston, VK;Sarisky, RT
通讯作者：
Sarisky, RT
影响因子：
7.3
作者：
Halgren, TA;Murphy, RB;Banks, JL
通讯作者：
Banks, JL
影响因子：
2.7
作者：
da Silva, AJM;Melo, PA;Costa, PRR
通讯作者：
Costa, PRR